Lower Protein Levels Research Articles

The challenges to detect, quantify, and characterize viral reservoirs in the current antiretroviral era

Abstract A major barrier to cure HIV is the early generation of viral reservoirs in tissues. These viral reservoirs can contain intact or defective proviruses, but both generates low levels of viral proteins contribute to chronic bystander damage even in the ART era. Most viral reservoir detection techniques are limited to blood-based, reactivation, and sequencing assays that lack spatial properties to examine the contribution of the host’s microenvironment to latency and cure efforts. Currently, little is known about the contribution of the microenvironment to viral reservoir survival, residual viral expression, and associated inflammation. Only a few spatiotemporal techniques are available, and fewer integrate spatial genomics, transcriptomics, and proteomics into the analysis of the viral reservoir microenvironment-all essential components to cure HIV. During the development of these spatial techniques, many considerations need to be included in the analysis to avoid misinterpretation. This manuscript tries to clarify some critical concepts in viral reservoir detection by spatial techniques and the upcoming opportunities for cure efforts.

UV Sensitivities of Two Littoral and Two Deep-Freshwater Amphipods (Amphipoda, Crustacea) Reflect Their Preferred Depths in the Ancient Lake Baikal

Solar ultraviolet (UV) is among the most important ecological factors shaping the composition of biota on the planet’s surface, including the upper layers of waterbodies. Inhabitants of dark environments recently evolving from surface organisms provide natural opportunities to study the evolutionary losses of UV adaptation mechanisms and better understand how those mechanisms function at the biochemical level. The ancient Lake Baikal is the only freshwater reservoir where deep-water fauna emerged, and its diverse endemic amphipods (Amphipoda, Crustacea) now inhabit the whole range from highly transparent littoral to dark depths of over 1600 m, which makes them a convenient model to study UV adaptation. With 10-day-long laboratory exposures, we show that adults of deep-water Baikal amphipods Ommatogammarus flavus and O. albinus indeed have high sensitivity to environmentally relevant UV levels in contrast to littoral species Eulimnogammarus cyaneus and E. verrucosus. The UV intolerance was more pronounced in deeper-dwelling O. albinus and was partially explainable by lower levels of carotenoids and carotenoid-binding proteins. Signs of oxidative stress were not found but UV-B specifically seemingly led to the accumulation of toxic compounds. Overall, the obtained results demonstrate that UV is an important factor limiting the distribution of deep-water amphipods into the littoral zone of Lake Baikal.

Understanding the Relationship between the Ketogenic Diet and the Heart: A Novel Therapeutic Potential for Cardiovascular Health.

Obesity and cardiovascular diseases are major health problems worldwide, and weight loss is used as a treatment strategy to enhance various aspects. While there are many weight loss methods, one of the most effective is through a dietary approach. The ketogenic diet (KD), which is characterized by low carbohydrates and high levels of fat and/or protein, is used in obese patients as it is a promising treatment option for weight loss as well as for controlling the risk factors for cardiovascular diseases, as seen in its effects on cardio-metabolic outcomes, particularly in obesity, heart failure, and hypertension. In this review, we summarize the clinical evidence of the efficacy and safety of the KD in controlling risk factors for cardiovascular diseases and discuss the possible mechanisms of action based on recent evidence in understanding the influence of the KD at the cellular and molecular levels.

Knockout of RIG-I in HEK293 cells by CRISPR/Cas9

We knocked out the retinoic acid-inducible gene I (RIG-I) in HEK293 cells via CRISPR/Cas9 to reveal the effects of RIG-I knockout on the key factors in the type I interferon signaling pathway. Three single guide RNAs (sgRNAs) targeting RIG-I were designed, and the recombination vectors were constructed on the basis of the pX459 vector and used to transfect HEK293 cells, which were screened by puromycin subsequently. Furthermore, a mimic of virus, poly I: C, was used to transfect the cells screened out. RIG-I knockout was checked by sequencing, real-time quantitative PCR, Western blotting, and immunofluorescence assay. Meanwhile, the expression levels of key factors of type I interferon signaling pathway such as melanoma differentiation-associated gene 5 (MDA5), interferonβ1 (IFNβ1), and nuclear factor-kappa B p65 [NF-κB(p65)], as well as cell viability, were determined. The results showed that two HEK293 cell lines (S1 and S3) with RIG-I knockout were obtained, which exhibited lower mRNA and protein levels of RIG-I than the wild type HEK293 cells (P < 0.05). The mRNA levels of MDA5 and IFNβ1 in S1 and S3 cells and the protein level of NF-κB(p65) in S3 cells were lower than those in the wild type (P < 0.05). More extranuclear NF-κB(p65) protein was detected in S1 cells than in the wild type after transfection with poly I: C. Plus, the wild-type and S1 cells transfected with poly I: C for 48 h showcased reduced viability (P < 0.05), while S3 cells did not display the reduction in cell viability. In summary, the present study obtained two HEK293 cell lines with RIG-I knockout via CRISPR/Cas9, which provided a stable cell model for exploring the mechanism of type I interferon signaling pathway.

Creating of novel Wx allelic variations significantly altering Wx expression and rice eating and cooking quality

Granule-bound starch synthase I (GBSSI) encoding gene Waxy (Wx), which largely regulates the amylose content of rice grains, is a master module determining rice eating and cooking quality (ECQ). Fine-tuning amylose level of grains is an ideal strategy to improve rice quality. Through fine editing of Wxa promoter and 5′UTR by CRISPR/Cas9 system, we created 14 types of novel Wx allelic variations, of which MT7 and MT13 were able to alter Wx expression and amylose content of grains. MT7 showed fragment deletion and base insertions in CAAT-boxes, hardly detectable expression levels of GBSSI mRNA and protein, and generated 5.87% amylose in grains. MT13 had fragment deletions in the A-box and the TATA-box, low expression levels of GBSSI mRNA and protein, and generated 9.61% amylose in grains. Besides of the amylose content, MT7 and MT13 significantly reduced protein content and increased lipid content of grains compared with Wxa. A comparison of MT7, MT13 and other allelic lines demonstrated the importance of base insertion around the second CAAT-box and 31bp-deletion following the second TATA-box in modulating Wx expression. Thus, our study generated two novel Wx allelic variations which significantly alter Wx expression and amylose content of rice grains, providing not only new germplasms for soft rice breeding, but also insights into candidate cis elements of Wx.

Integration of transcriptome, histopathology, and physiological indicators reveals regulatory mechanisms of largemouth bass (Micropterus salmoides) in response to carbonate alkalinity stress

The use of saline-alkali water resources is an important initiative for freshwater aquaculture. Largemouth bass (Micropterus salmoides) shows some salinity tolerance, but little is known about its alkali adaptation mechanism. First, we determined the 96-h 50 % lethal dose of alkaline carbonate (96-h LC50) for largemouth bass (41.41 mmol/L). Then, largemouth bass were subjected to 96-h carbonate alkalinity treatments and their responses were determined on the basis of histopathological, biochemical, and transcriptome analyses. The experiment included a control group (CA0, 0.49 mmol/L) and two treatment groups (CA20, 20 mmol/L; CA45, 45 mmol/L). The gill and liver tissues of the CA45 group exhibited severe deformation, including a thickened matrix, gill cell detachment, hepatocyte swelling, increased vacuolization, and localized necrotic lesions. In the CA20 group, some gill filaments were curled and deformed, hepatocytes had migrated, and tissue damage was minor. Both groups showed a lower percentage of apoptotic cells in gill and liver tissues. Serum antioxidant capacity first increased and then decreased, whereas energy metabolism and hormone indexes significantly increased under alkaline carbonate stress. Liver injury in the CA45 group resulted in significantly lower levels of cortisol, lactate dehydrogenase, and total protein compared with those in the CA20 group (P < 0.05). Transcriptome analyses of gill tissues from the CA45 group revealed significant alterations in pathways related to steroid biosynthesis (sqlea, lss, msmo1, tm7sf2), terpenoid backbone biosynthesis (hmgcs1, fdps, hmgcra, mvda), and ECM-receptor interaction and focal adhesion (col1a, col2a1b, col9a, col6a) due to alkalinity stress. These results indicate that liver damage under high alkalinity stress inhibits energy supply, while gill cell dysregulation triggers gill lesions.

Whole-exome sequencing reveals genomic landscape of intrahepatic cholangiocarcinoma and identifies SAV1 as a potential driver.

Intrahepatic cholangiocarcinoma (ICC) is the second most common primary hepatic malignancy after hepatocellular carcinoma, with poor prognosis and limited treatment options. The genomic features of ICC in Chinese patients remain largely unknown. In this study, we perform deep whole-exome sequencing of 204 Chinese primary ICCs and characterize genomic alterations and clonal evolution, and reveal their associations with patient outcomes. We identify six mutational signatures, including Signatures A and F, which are highly similar to previously described signatures linked to aristolochic acid and aflatoxin exposures, respectively. We also identify 13 significantly mutated genes in the ICC samples, including SAV1. We find that SAV1 was mutated in 2.9% (20/672) of 672 ICC samples. SAV1 mutation is associated with lower SAV1 protein levels, higher rates of tumor recurrence, and shorter overall patient survival. Biofunctional investigations reveal a tumor-suppressor role of SAV1: its inactivation suppresses Hippo signaling, leading to YAP activation, thereby promoting tumor growth and metastasis. Collectively, our results delineate the genomic landscape of Chinese ICCs and identify SAV1 as a potential driver of ICC.

Effects of adding niacinamide to diets with normal and low protein levels on the immunity, antioxidant, and intestinal microbiota in growing-finishing pigs

This study aimed to investigate the effects of nicotinamide (NAM) applied to diets with different crude protein levels on immune function, antioxidant capacity, and intestinal flora in growing-finishing pigs. Forty barrows (37.0±1.0 kg) were randomly allocated to one of four dietary treatments (n=10 per group). The diets in the two phases consisted of a basal diet with 30 mg/kg NAM, a basal diet with 360 mg/kg NAM, a low-protein diet with 30 mg/kg NAM, and a low-protein diet with 360 mg/kg NAM. The results showed that dietary addition of 360 mg/kg NAM decreased IL-12, malondialdehyde, IgG and IgM contents in the plasma and increased total superoxide dismutase activity and total antioxidant capacity in the colonic mucosa (P < .05). Supplementing the diet with 360 mg/kg NAM increased mRNA expression of the nucleotide-binding oligomerization domain containing 2 and nuclear factor erythroid 2-related factor 2 and protein expression of nuclear factor kappa-B and toll-like receptor 4 in the colonic mucosa (P < .05). The concentrations of acetic acid and butyric acid in the colonic contents and the abundance of Actinobacteriota in the colon at the phylum level were significantly decreased by feeding low-protein diets (P < .05). Additionally, the addition of 360 mg/kg NAM to diets increased (P < .05) the Sobs, Ace, and Chao indices of colonic microorganisms in pigs. Overall, the rational use of NAM can improve inflammatory status, enhance antioxidant capacity and intestinal barrier function, and increase colonic microbial diversity in growing-finishing pigs.

Orf116b Induces Pollen Abortion in a Novel Cotton (Gossypium hirsutum L.) Cytoplasmic Male Sterile Line J4A.

Cytoplasmic male sterile (CMS) systems have great potential in hybrid seed production in many plants. However, the incompatibility between the cytoplasmic and nuclear genes and the availability of fewer CMS lines limit the system's application in cotton heterosis. Therefore, the present study sequenced the mitochondrial (mt) genomes of a novel cotton (Gossypium hirsutum L.) CMS line J4A and its cytoplasmic nuclear homologous maintainer line J4B to investigate the mechanism underlying CMS and improve its application. A novel CMS gene, orf116b, was identified in the CMS line. Ectopic expression of orf116b in Escherichia coli suppressed growth, while its overexpression in Arabidopsis, rice, tobacco, and cotton led to complete or partial male sterility. Further analysis of anthers revealed mt disruption in J4A with higher levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA), but lower levels of ATP and ribosomal protein (RP) than in J4B. Finally, a membrane-based yeast two-hybrid (MYTH) assay and bimolecular fluorescence complementation (BiFC) assays demonstrated that orf116b probably interacts with an anther-specific protein, LAT52. These observations collectively proved that orf116b is associated with early and stable pollen abortion in cotton, providing a foundation for further research on cotton fertility restoration and heterosis breeding.

Impact of tiger nut milk as a substitute for cow milk on the rheological, physiochemical, and organoleptic properties of functional ice cream

AbstractTiger nuts are an excellent food choice due to their rich mineral, fiber, and antioxidant content. Tiger nut milk was replaced by 25, 50, 75, and 100% of cow’s milk in the ice cream mixture. The impact of substituting cow milk with tiger nut milk on the physicochemical, rheological, and organoleptic properties of ice cream was examined. Adding tiger nut milk in a combination resulted ice cream mixes with more fiber and antioxidants and a lower protein and ash levels compared to the control (100% cow milk). Ice cream with tiger nut milk was higher in Fe and Zn than that of control. Dynamic viscosity values of ice cream mixes with tiger nut milk were decreased with increasing values of the applied shear rate compared to the control. All functional ice cream formulations with tiger nut milk showed higher specific gravity and melting rate with lower overrun percentages than the control. Regarding taste and flavor, structure, texture, and melting in the mouth, there is no appreciable difference between the T25, T50, and control treatments. Therefore, cow’s milk can be replaced with tiger nut milk up to 50% in the ice cream industry to raise the nutritional value and functional properties.

Abstract 4131381: Comparison of Dietary Macronutrient Interventions for Weight and Cardiovascular Risk Factor Reduction: A Systematic Review and Network Meta-analysis of Randomized Controlled Trial

Background: Dietary interventions play a crucial role in weight management and reducing cardiovascular risk factors. Our study aims to compare the effectiveness of four dietary macronutrient interventions on weight loss and cardiovascular (CV) risk factor reduction through a systematic review and network meta-analysis. Methods: We conducted a comprehensive literature search on PubMed, Scopus, Embase, and Cochrane Library up till May 2024 to identify randomized controlled trials (RCTs) comparing four macronutrient dietary interventions including Mediterranean Diet (MD), Keto, Dietary Approaches to Stop Hypertension (DASH), and Intermittent Fasting (IF) with study period ≥ 6 months or 24 weeks. The primary outcomes of interest were weight loss, systolic blood pressure (SBP), Diastolic blood pressure (DBP), Body Mass Index (BMI), High density lipoprotein (HDL), Low density Lipoprotein (LDL), cholesterol levels and C-reactive protein (CRP) levels. Outcomes were reported as standard mean difference (SMD). Results: Our analysis identified 50 studies enrolling 5368 patients (MD=3554; DASH=838; Keto=206; IF=770). Regarding BP outcome, MD and DASH had significant reduction in SBP and DBP respectively (MD [SBP]: -0.76 mmHg vs DASH [DBP]: -1.92 mmHg) respectively. In contrast, IF showed a significant rise in SBP (0.87). MD participants also had significant weight loss (-1.06 kg) and a moderate decrease in BMI (-0.79) when compared with other diets. Furthermore, IF, keto, and MD showed moderate increase in HDL levels (0.61, 0.77 and 0.33) respectively. In contrast, DASH resulted in a moderate decline in HDL levels (-0.92). IF and MD resulted in modest decline in LDL levels (-0.45 and -0.42) respectively. In contrast, Keto demonstrated non-significant rise in LDL (0.35). DASH showed a significant decrease in triglycerides (-3.02). Lastly, MD demonstrated a significant reduction in CRP (-0.89). Conclusions: MD and DASH were superior to other dietary interventions in terms of weight loss and CV risk factors. Further research is required to tailor specific types of dietary interventions and assess their long-term efficacy on weight loss and CV risk reduction.

Evaluating sustainable energy pathways: Economic perspective on advanced hydrogen production

This investigation seeks to evaluate hydrogen generation technologies in China with a focus on two commercial processes, namely hydrothermal oxidation and supercritical water gasification of organic waste from 2011 to 2021. This period witnessing major development, and highlight the key determinants of hydrogen production. The present study was mainly concerned with the enhancement of SCWG processes for hydrogen generation from microalgae and other potential feedstock. Some of the significant observations made in this period are the critical role of biomass composition, choice of catalyst, and operating conditions in increasing H-producing capabilities of reactor systems. The authors of research affirm that the feedstock with a high carbohydrate content and low levels of protein are the most suitable for hydrogen generation. Thus, expanding research on Supercritical Water Gasification, there is the development of HTO methods to consider various pathways of hydrogen production and their connection with existing energy networks. It also studied the effects that several types of catalysts embracing the heterogeneous transition metals and the homogeneous alkali metal compounds have on the efficiency of hydrogen production. These changes have rendered the hydrogen generation process to make more precise predictions and/or course corrections on the process. The studies that have been done between 2011 and 2021 show significant improvements in the ways that sustainable and efficient hydrogen generation technology can be created in China. The results provide a solid platform for future improvements and stress positive impacts that can be provided by these technologies for China's sustainability and energy transition progress.

BIOM-47. SINGLE NUCLEOTIDE POLYMORPHISM IN SMARCB1 MUTATION FOR SCHWANNOMATOSIS

Abstract Schwannomatosis, a form of neurofibromatosis, involves critical roles for the SMARCB1 gene. Clinical diagnosis guidelines recommend considering pathogenic variants or suspecting schwannomatosis to exclude splicing-related intron mutations. However, some patients may initially present with non-significant mutations and lack clinical signs of schwannoma, leading to misdiagnosis, missed annual assessments, malignancy awareness, and genetic consultation opportunities during pregnancy. Here, we reported a patient with a sporadic synonymous mutation in the SMARCB1 gene (SNP c.1032 C&amp;gt;T (p.Gly344Gly(GGC&amp;gt;GGT)) in exon 8), not previously associated with schwannomatosis. This patient, a 55-year-old female with an 8-year history of schwannomatosis, presented to the neurosurgical department for recurrent tumor removal. Initially, the clinical impact of the SMARCB1 mutation was undetermined, and other genes tested were wild-type. Both schwannomas exhibited a mosaic loss of nuclear SMARCB1 protein, ranging from 10% to 60%. Protein assays confirmed low levels of SMARCB1 protein, with a significant loss in the distal thigh schwannoma. It highlights the tumorigenic potential of single nucleotide polymorphism in the SMARCB1 gene and aids clinicians in making more accurate diagnoses in patients with pathologically confirmed schwannoma. It underscores the importance of long-term follow-up, increased awareness of recurrence and malignancy, and timely surgical planning in such patients.

CTNI-45. PHASE 2 STUDY OF SORAFENIB, VALPROIC ACID, AND SILDENAFIL IN THE TREATMENT OF RECURRENT HIGH-GRADE GLIOMA

Abstract PURPOSE Here we report the results of a single-center phase 2 clinical trial combining sorafenib tosylate, valproic acid, and sildenafil for the treatment of patients with recurrent high-grade glioma. PATIENTS AND METHODS Eligible patients were 18 years of age and older with an Eastern Cooperative Oncology Group performance status of 0-2 and pathologically confirmed high-grade glioma (WHO grade 3 or 4), with documented CT or MRI progression or recurrence and measurable or evaluable disease. The treatment schedule was a combination of sorafenib, valproic acid, and sildenafil, each agent administered orally, twice daily continuously. A cycle consisted of 4 weeks. RESULTS Clinical toxicities were grade 1 and grade 2, with one grade 3 toxicity for maculopapular rash (6.4%). For all evaluable patients, the median progression-free survival was 3.65 months and overall survival (OS) 10.0 months. There was promising evidence showing clinical activity and benefit. In the 33 evaluable patients, low protein levels of the chaperone GRP78 (HSPA5) was significantly associated with a better OS (p &amp;lt; 0.0026). A correlation between the expression of PDGFRa and OS approached significance (p &amp;lt; 0.0728). Five patients presently have a mean OS of 73.6 months and remain alive. CONCLUSIONS This is the first therapeutic intervention glioblastoma trial to significantly associate GRP78 expression to OS. Our data suggest that the combination of sorafenib tosylate, valproic acid, and sildenafil requires additional clinical development in the recurrent glioma population.

An infant case of coronary artery aneurysms with no systemic symptoms after treatment for Kawasaki disease.

This case is a 4-month-old patient with Kawasaki disease who showed coronary artery lesions after a disease relapse. Following the guidelines from the Randomised Controlled Trial to Assess Immunoglobulin plus Steroid Efficacy, the patient initially saw an improvement in symptoms. However, a relapse occurred despite low levels of C-reactive protein, leading to significant coronary artery growth. Treatment with a third dose of intravenous immunoglobulin and cyclosporine led to a slow reduction in the size of the coronary artery abnormalities. This case highlights the difficulty in monitoring Kawasaki disease progression through clinical symptoms and C-reactive protein levels alone. We also emphasise the necessity of echocardiographic monitoring in patients receiving anti-inflammatory treatments, including steroids, because coronary artery problems can arise or continue without the usual signs of Kawasaki disease or increased C-reactive protein levels.

SlMIPS2, a myo-inositol phosphate synthase gene, regulates phosphate homeostasis by influencing SlPHL1 and SlSPX2 levels in tomato seedlings.

Phosphorus (P) is a quintessential macronutrient utilized by plants to support various metabolic processes during growth and development. Recent studies have revealed the pivotal role of inositol hexa-kis/pyrophosphate (InsP6-8), the derivatives of Myo-inositol (MI), in facilitating the interaction between SYG1/PHO81/XPR1 (SPX) and Phosphate starvation response (PHR) proteins. Myo-inositol phosphate synthase (MIPS) catalyzes the first committed step in MI biosynthesis. Although the role of MIPS genes in mediating stress responses in plants is well elucidated, its role in phosphate (Pi) deficiency remains largely unexplored. This study demonstrates that out of the five MIPS genes encoded by the tomato genome, only SlMIPS2 is sharply induced at an early stage of Pi starvation in tomato seedlings. Silencing of SlMIPS2 led to improved seedling growth with enhanced total soluble Pi and total P levels in the silenced plants under high Pi availability. SlMIPS2 silencing also caused a significant reduction in MI and InsP6 content in the tomato seedlings. These seedlings with depleted InsP6 levels accumulated lower levels of SlSPX2 protein. In contrast, stabilized SlPHL1 levels were noticed in these plants, directly implicating this transcription factor in activating phosphate starvation inducible (PSI) genes in the SlMIPS2-silenced seedlings, even under high Pi conditions. The results assign a novel role to SlMIPS2 in regulating cellular InsP6 levels and SPX-PHR interactions to control Pi homeostasis in tomato seedlings.

Panax notoginseng saponins improves lipid metabolism and prevents atherosclerosis in mice with steroid-resistant lupus nephritis via the SIRT1/PPARγ signaling pathway

Steroids serve as the primary medication for treating lupus nephritis (LN), however, steroid-resistance (SR) occurs sporadically in clinical practice, significantly affecting the therapeutic effect and long-term prognosis of patients. Our previous study found that panax notoginseng saponins (PNS) could partially reverse SR in LN. To further explore the role of PNS in reversing SR and reducing cardiovascular complications in LN, we conducted this study. Lupus mice were induced into SR while simultaneously receiving PNS. SIRT1-siRNA, SIRT1-siRNA NC, normal and lupus mice were used as control groups. Urine protein levels were measured at week 0, 4 and 8. Lipid metabolism-related biomarkers and renal function were assessed. The apoptosis rate of abdominal aortic endothelial cells was detected using flow-cytometry. The expression levels of PPARγ and SIRT1 were measured using RT-PCR and Western Blotting. Immunohistochemistry was performed to examine ACAT1 and VCAM-1 expressions. The results showed that compared to the SR lupus mice, the lupus mice treated with low/high dose PNS presented lower levels of urinary protein, serum creatinine, and blood lipids, a lower apoptosis rate of abdominal aortic endothelial cells, and decreased levels of ACAT1 and VCAM-1 PI in liver tissue, while the high-dose PNS exhibited more evidently. The PPARγ expression in SIRT1-siRNA group, as well as in low-dose and high-dose PNS groups was higher than that in the lupus and SR lupus group. In contrast, the expression of SIRT1 showed the opposite trend. Therefore, we conclude that PNS has the efficacy of reversing SR and ameliorating dyslipidemia in LN by modulating the SIRT1/PPARγ signaling pathway.

The application effect of autologous platelet-rich plasma combined with negative pressure sealing drainage technology in pressure ulcer wound repair.

This study aimed to evaluate the efficacy of combining autologous platelet-rich plasma (PRP) with negative pressure wound therapy (NPWT) for the repair of pressure ulcers. We included 90 patients with pressure ulcers from General Technology Gemstone Flower Healthcare Jilin City Hospital of Chemical Industry between January 2021 and December 2023. Patients were randomly assigned to either a control group or an observation group, with 45 patients in each. The control group received NPWT alone, while the observation group received a combination of PRP and NPWT. Outcomes were compared between the groups, including clinical efficacy, wound recovery time (infection control time, wound healing time, hospital stay), levels of inflammatory markers (C-reactive protein, erythrocyte sedimentation rate, white blood cell count), pain scores (numerical rating scale), Pressure Ulcer Scale for Healing scores, and incidence of complications. The observation group demonstrated a total effective rate of 95.56% (43/45), significantly higher than the control group's 80.00% (35/45) (P < .05). Additionally, the observation group had significantly shorter infection control times, wound healing times, and hospital stays (P < .05). Two weeks post-surgery, the observation group had significantly lower levels of C-reactive protein, erythrocyte sedimentation rate, and white blood cell count, as well as reduced numerical rating scale and Pressure Ulcer Scale for Healing scores (P < .05). The incidence of complications was 6.67% (3/45) in the observation group compared to 33.33% (15/45) in the control group, with a significant difference (P < .05). Combining autologous PRP with NPWT significantly improves clinical outcomes, reduces inflammatory responses, decreases pain, accelerates wound healing, and lowers complication rates in patients with pressure ulcers.

A sort and sequence approach to dissect heterogeneity of response to a self-amplifying RNA vector in a novel human muscle cell line

Self-amplifying RNA (saRNA) is an extremely promising platform because it can potentially produce more protein for less RNA. We used a ‘sort and sequence’ approach to identify host cell factors associated with transgene expression from saRNA; the hypothesis was that cells with different expression levels would have different transcriptomes. We tested this in CDK4/hTERT immortalised human muscle cells transfected with VEEV derived saRNA encoding GFP. Cells with the highest expression levels had very high levels of transgene mRNA (5-10% total reads); the cells sorted with low and negative levels of GFP protein also had detectable levels of both VEEV and GFP RNA. To understand host responses, we performed RNASeq. Differentially expressed gene (DEG) patterns varied with GFP expression; GFP high cells, had many more DEG, these were associated with protein synthesis and cell metabolism. Comparing profiles by an unsupervised approach revealed that negative cells expressed higher levels of cell intrinsic immunity genes such as IFIT1, MX1, TLR3 and MyD88. To explore the role of interferon, cells were treated with the Jak inhibitor Ruxolitinib, this reduced DEG number but differences between cells sorted by expression level remained. These studies demonstrate the complex interplay of factors, some immune related, affecting saRNA transgenes.

Mitochondrial respiratory complex II is altered in renal carcinoma

BackgroundRenal cell carcinoma (RCC) is a disease typified by anomalies in cell metabolism. The function of mitochondria, including subunits of mitochondrial respiratory complex II (CII), in particular SDHB, are often affected. Here we investigated the state and function of CII in RCC patients. MethodsWe evaluated tumour tissue as well as the adjacent healthy kidney tissue of 78 patients with RCC of different histotypes, focusing on their mitochondrial function. As clear cell RCC (ccRCC) is by far the most frequent histotype of RCC, we focused on these patients, which were grouped based on the pathological WHO/ISUP grading system to low- and high-grade patients, indicative of prognosis. We also evaluated mitochondrial function in organoids derived from tumour tissue of 7 patients. ResultsccRCC tumours were characterized by mutated von Hippel-Lindau gene and high expression of carbonic anhydrase IX. We found low levels of mitochondrial DNA, protein and function, together with CII function in ccRCC tumour tissue, but not in other RCC types and non-tumour tissues. Mitochondrial content increased in high-grade tumours, while the function of CII remained low. Tumour organoids from ccRCC patients recapitulated molecular characteristics of RCC tissue. ConclusionsOur findings suggest that the state of CII, epitomized by its assembly and SDHB levels, deteriorates with the progressive severity of ccRCC. These observations hold the potential for stratification of patients with worse prognosis and may guide the exploration of targeted therapeutic interventions.